Response of particle-attached and free-living bacterial communities to Microcystis blooms.

The massive proliferation of Microcystis threatens freshwater ecosystems and degrades water quality globally. Understanding the mechanisms that contribute to Microcystis growth is crucial for managing Microcystis blooms. The lifestyles of bacteria can be classified generally into two groups: particle-attached (PA; > 3 µm) and free-living (FL; 0.2-3.0 µm). However, little is known about the response of PA and FL bacteria to Microcystis blooms. Using 16S rRNA gene high-throughput sequencing, we investigated the stability, assembly process, and co-occurrence patterns of PA and FL bacterial communities during distinct bloom stages. PA bacteria were phylogenetically different from their FL counterparts. Microcystis blooms substantially influenced bacterial communities. The time decay relationship model revealed that Microcystis blooms might increase the stability of both PA and FL bacterial communities. A contrasting community assembly mechanism was observed between the PA and FL bacterial communities. Throughout Microcystis blooms, homogeneous selection was the major assembly process that impacted the PA bacterial community, whereas drift explained much of the turnover of the FL bacterial community. Both PA and FL bacterial communities could be separated into modules related to different phases of Microcystis blooms. Microcystis blooms altered the assembly process of PA and FL bacterial communities. PA bacterial community appeared to be more responsive to Microcystis blooms than FL bacteria. Decomposition of Microcystis blooms may enhance cooperation among bacteria. Our findings highlight the importance of studying bacterial lifestyles to understand their functions in regulating Microcystis blooms. KEY POINTS: • Microcystis blooms alter the assembly process of PA and FL bacterial communities • Microcystis blooms increase the stability of both PA and FL bacterial communities • PA bacteria seem to be more responsive to Microcystis blooms than FL bacteria.

How do freshwater microalgae and cyanobacteria respond to antibiotics?

Antibiotic pollution is an emerging environmental challenge. Residual antibiotics from various sources, including municipal and industrial wastewater, sewage discharges, and agricultural runoff, are continuously released into freshwater environments, turning them into reservoirs that contribute to the development and spread of antibiotic resistance. Thus, it is essential to understand the impacts of antibiotic residues on aquatic organisms, especially microalgae and cyanobacteria, due to their crucial roles as primary producers in the ecosystem. This review summarizes the effects of antibiotics on major biological processes in freshwater microalgae and cyanobacteria, including photosynthesis, oxidative stress, and the metabolism of macromolecules. Their adaptive mechanisms to antibiotics exposure, such as biodegradation, bioadsorption, and bioaccumulation, are also discussed. Moreover, this review highlights the important factors affecting the antibiotic removal pathways by these organisms, which will promote the use of microalgae-based technology for the removal of antibiotics. Finally, we offer some perspectives on the opportunities for further studies and applications.

Mucilaginibacter straminoryzae sp. nov., isolated from rice straw used for growing periphyton.

A rod-shaped, non-motile, Gram-negative bacterium, strain RS28T, was isolated from rice straw used as material for periphyton growth. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain RS28T was affiliated with the genus Mucilaginibacter and had the highest sequence similarity to Mucilaginibacter ginkgonis HMF7856T (96.47 %) and Mucilaginibacter polytrichastri DSM 26907T (96.12 %). Strain RS28T was found to grow at pH 5.5-8.0, 17-40 °C and in the presence of 0-1.5 % (w/v) NaCl. Strain RS28T contained summed feature 3 (comprising C16 : 1 ω7c and/or C16 : 1 ω6c), iso-C15 : 0 and iso-C17 : 0 3-OH as the major fatty acids (> 10.0 %). The major polar lipids were phosphatidylethanolamine, two unidentified phospholipids, two unidentified aminophospholipids, three unidentified aminolipids and one unidentified lipid. The respiratory quinone was menaquinone 7. The genomic DNA G+C content was 44.7 mol%. Strain RS28T possessed six putative secondary metabolite gene clusters involved in the synthesis of resorcinol, NRPS-like, terpene, lassopeptide, T3PKS and arylpolyene. On the basis of the phenotypic, chemotaxonomic, and phylogenetic characteristics, strain RS28T represents a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter straminoryzae sp. nov. is proposed. The type strain is RS28T (=KCTC 92039T=LMG 32424T).

Lacibacter sediminis sp. nov., isolated from contaminated freshwater sediment.

Two Gram-stain-negative bacterial strains, S13-6-6 and S13-6-22T, were isolated from sediment sample collected at a water depth of 4 m from Lake Hongze, Jiangsu Province, PR China. The cells of strains S13-6-6 and S13-6-22T were non-spore-forming, aerobic, non-motile and formed orange colonies on R2A agar. Comparative 16S rRNA gene sequence studies revealed a clear affiliation of the two strains with he phylum Bacteroidota, and revealed the highest pairwise sequence similarities with Lacibacter daechungensis H32-4T (97.8 %), Lacibacter cauensis NJ-8T (97.8 %), Lacibacter luteus TTM-7T (97.4 %) and Lacibacter nakdongensis SS2-56T (97.4 %). The results of phylogenetic analysis based on 16S rRNA gene sequences indicated that the strains formed a clear phylogenetic lineage with the genus Lacibacter. The major fatty acids were identified as iso-C15 : 1G, iso-C15 : 0, iso-C17 : 0 3-OH and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) (>10 %), and the respiratory quinone was identified as menaquinone MK-7. The polar lipids consisted of phosphatidylethanolamine, two unidentified aminolipids, an unidentified phospholipid and six unidentified lipids. The genomic DNA G+C content was determined to be 40.2 mol% (HPLC) for strain S13-6-6 and 40.3 % (genome) for strain S13-6-22T. The combined genotypic and phenotypic data indicated that strains S13-6-6 and S13-6-22T represent a novel species of the genus Lacibacter, for which the name Lacibacter sediminis sp. nov. is proposed. The type strain is S13-6-22T (=CGMCC 1.17450T =JCM 35802T).

Novosphingobium cyanobacteriorum sp. nov., isolated from a eutrophic reservoir during the Microcystis bloom period.

A novel Gram-stain-negative, aerobic and rod-shaped bacterial strain, HBC54T, was isolated from periphyton during a Microcystis bloom. Based on the results of the 16S rRNA gene sequence analysis, strain HBC54T was closely related to Novosphingobium aerophilum 4Y4T (98.36 %), Novosphingobium aromaticivorans DSM 12444T (98.08 %), Novosphingobium huizhouense c7T (97.94 %), Novosphingobium percolationis c1T (97.65 %), Novosphingobium subterraneum DSM 12447T (97.58 %), Novosphingobium olei TW-4T (97.58 %) and Novosphingobium flavum UCT-28T (97.37 %). The average nucleotide identity and digital DNA-DNA hybridization values between HBC54T and its related type stains were below 78.97 and 23.7 %, which are lower than the threshold values for species delineation. The major fatty acids (>10.0 %) were identified as C14 : 0 2-OH, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and the respiratory quinone was ubiquinone Q-10. The main polar lipids detected in the strain were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, diphosphatidylglycerol and three unidentified phospholipids. The genomic DNA G+C content was 64.8 mol%. Strain HBC54T is considered to represent a novel species within the genus Novosphingobium, for which the name Novosphingobium cyanobacteriorum sp. nov. is proposed. The type strain is HBC54T (=KCTC 92033T=LMG 32427T).

Description of Hymenobacter sediminicola sp. nov., isolated from contaminated sediment.

A polyphasic taxonomic study was conducted on two Gram-negative, non-sporulating, non-motile bacterial strains, S2-20-2T and S2-21-1, isolated from a contaminated freshwater sediment in China. Comparative 16S rRNA gene sequence studies revealed a clear affiliation of two strains with Bacteroidetes, which showed the highest pairwise sequence similarities with Hymenobacter duratus BT646T (99.3%), Hymenobacter psychrotolerans Tibet-IIU11T (99.3%), Hymenobacter kanuolensis T-3T (97.6%), Hymenobacter swuensis DY53T (96.9%), Hymenobacter tenuis POB6T (96.8%), Hymenobacter seoulensis 16F7GT (96.7%), and Hymenobacter rigui KCTC 12533T (96.5%). The phylogenetic analysis based on 16S rRNA gene sequences showed that two strains formed a clear phylogenetic lineage with the genus Hymenobacter. Major fatty acids were identified as iso-C15:0, anteiso-C15:0, and summed feature 3 (C16:1 ω6c and/or C16:1 ω7c/t) and summed feature 4 (iso-C17:1 I and/or anteiso-C17:1 B). Major cellular polar lipids were identified as phosphatidylethanolamine, three unidentified aminolipids, an unidentified aminophosopholipid and an unidentified lipid. The respiratory quinone was detected as MK-7 and the genomic DNA G + C content was determined to be 57.9% (genome) for type strain S2-20-2T and 57.7 mol% (HPLC) for strain S2-21-1. The observed ANI and dDDH values between strain S2-20-2T and its closely related strains were 75.7-91.4% and 21.2-43.9%, respectively. Based on physiological, biochemical, genetic and genomic characteristics, we propose that strains S2-20-2T and S2-21-1 represent a novel species of the genus Hymenobacter, for which the name Hymenobacter sediminicola sp. nov. is proposed. The type strain is S2-20-2T (= CGMCC 1.18734T = JCM 35801T).

Hymenobacter cyanobacteriorum sp. nov., isolated from a freshwater reservoir during the cyanobacterial bloom period.

A Gram-negative, red-colored, and rod-shaped bacterial strain, DH14T, was isolated from a eutrophic reservoir. The 16S rRNA gene sequence analysis showed that strain DH14T was most closely related to Hymenobacter terrigena (98.3% similarity) and Hymenobacter terrae (98.1%). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain DH14T and its related type strains were below 82.9% and 27.2%, respectively. Strain DH14T contained iso-C15:0 (32.6%), anteiso-C15:0 (14.0%), and summed feature 3 (C16:1 ω6c and/or C16:1 ω7c) (25.8%) as major cellular fatty acids. The main polar lipids were phosphatidylethanolamine, two unidentified aminophospholipids, and one unidentified lipid. The respiratory quinone was menaquinone 7 (MK-7). The genomic DNA G + C content was 62.1%. These evidences support the classification of strain DH14T as a novel species in the genus Hymenobacter, for which the name Hymenobacter cyanobacteriorum sp. nov. is proposed. The type strain is DH14T (= KCTC 92040T = LMG 32425T).

Panacibacter microcysteis sp. nov., isolated from a eutrophic reservoir during the Microcystis bloom period.

A Gram-stain-negative, rod-shaped bacterial strain DH6T was isolated from fresh water of the Daechung Reservoir during the Microcystis bloom period. The strain grew at pH 6.0-8.5, at temperature 17-40 °C, and at 0-1% (w/v) NaCl concentration. Comparison of 16S rRNA gene sequence indicated that strain DH6T exhibits the highest similarity with Panacibacter ginsenosidivorans Gsoil 1550T (96.6%). The average nucleotide identity (ANI), digital DNA-DNA hybridization (dDDH), and average amino acid identity (AAI) values of strain DH6T compared to its related type strains were below 74.2%, 22.3%, and 74.8%, respectively. The predominant fatty acids (> 5.0%) were identified as iso-C17:0 3-OH, iso-C13:0, iso-C15:0, C17:0 2-OH, iso-C11:0, anteiso-C13:0, and iso-C15:1 G. The polar lipid profile contained phosphatidylethanolamine, four unidentified aminolipids, and three unidentified lipids. The respiratory quinone was menaquinone 7 (MK-7). The genomic DNA G + C content was 42.6%. Collectively, strain DH6T should be classified as a novel species within the genus Panacibacter, for which the name Panacibacter microcysteis sp. nov. is proposed. The type strain is DH6T (= KCTC 82471T = LMG 32426T).

Development and Applications of CRISPR/Cas9-Based Genome Editing in Lactobacillus.

Lactobacillus, a genus of lactic acid bacteria, plays a crucial function in food production preservation, and probiotics. It is particularly important to develop new Lactobacillus strains with superior performance by gene editing. Currently, the identification of its functional genes and the mining of excellent functional genes mainly rely on the traditional gene homologous recombination technology. CRISPR/Cas9-based genome editing is a rapidly developing technology in recent years. It has been widely applied in mammalian cells, plants, yeast, and other eukaryotes, but less in prokaryotes, especially Lactobacillus. Compared with the traditional strain improvement methods, CRISPR/Cas9-based genome editing can greatly improve the accuracy of Lactobacillus target sites and achieve traceless genome modification. The strains obtained by this technology may even be more efficient than the traditional random mutation methods. This review examines the application and current issues of CRISPR/Cas9-based genome editing in Lactobacillus, as well as the development trend of CRISPR/Cas9-based genome editing in Lactobacillus. In addition, the fundamental mechanisms of CRISPR/Cas9-based genome editing are also presented and summarized.

New Multidrug Efflux Systems in a Microcystin-Degrading Bacterium Blastomonas fulva and Its Genomic Feature.

A microcystin-degrading bacterial strain, Blastomonas fulva T2, was isolated from the culture of a microalgae Microcystis. The strain B. fulva T2 is Gram-stain-negative, non-motile, aerobic, non-spore-forming and phototrophic. The cells of B. fulva T2 are able to grow in ranges of temperature from 15 to 37 °C, with a pH of 6 to 8 and a salinity of 0 to 1% NaCl. Here, we sequenced the complete genome of B. fulva T2, aiming to better understand the evolutionary biology and the function of the genus Blastomonas at the molecular level. The complete genome of B. fulva T2 contained a circular chromosome (3,977,381 bp) with 64.3% GC content and a sizable plasmid (145.829 bp) with 60.7% GC content which comprises about 3.5% of the total genetic content. A total of 3842 coding genes, including 46 tRNAs and 6 rRNAs, were predicted in the genome. The genome contains genes for glycolysis, citric acid cycle, Entner-Doudoroff pathways, photoreaction center and bacteriochlorophylla synthesis. A 7.9 K gene cluster containing mlrA, mlrB, mlrC and mlrD1,2,3,4 of microcystin-degrading enzymes was identified. Notably, eight different efflux pumps categorized into RND, ABC and MFS types have been identified in the genome of strain T2. Our findings should provide new insights of the alternative reaction pathway as well as the enzymes which mediated the degradation of microcystin by bacteria, as well as the evolution, architectures, chemical mechanisms and physiological roles of the new bacterial multidrug efflux system.

The NanDeSyn database for Nannochloropsis systems and synthetic biology.

Nannochloropsis species, unicellular industrial oleaginous microalgae, are model organisms for microalgal systems and synthetic biology. To facilitate community-based annotation and mining of the rapidly accumulating functional genomics resources, we have initiated an international consortium and present a comprehensive multi-omics resource database named Nannochloropsis Design and Synthesis (NanDeSyn; http://nandesyn.single-cell.cn). Via the Tripal toolkit, it features user-friendly interfaces hosting genomic resources with gene annotations and transcriptomic and proteomic data for six Nannochloropsis species, including two updated genomes of Nannochloropsis oceanica IMET1 and Nannochloropsis salina CCMP1776. Toolboxes for search, Blast, synteny view, enrichment analysis, metabolic pathway analysis, a genome browser, etc. are also included. In addition, functional validation of genes is indicated based on phenotypes of mutants and relevant bibliography. Furthermore, epigenomic resources are also incorporated, especially for sequencing of small RNAs including microRNAs and circular RNAs. Such comprehensive and integrated landscapes of Nannochloropsis genomics and epigenomics will promote and accelerate community efforts in systems and synthetic biology of these industrially important microalgae.

Cochlodiniinecator piscidefendens gen. nov., sp. nov., an algicidal bacterium against the ichthyotoxic dinoflagellate Cochlodinium polykrikoides.

Harmful algal blooms caused by Cochlodinium polykrikoides result in enormous economic damage to the aquaculture industry. Biological control methods have attracted wide attention due to their environmental-friendliness. In this study, a novel algicidal bacterium, designated strain M26A2MT, was determined for its taxonomic position and was evaluated for its potential to mitigate C. polykrikoides blooms. Strain M26A2MT exhibited the highest 16S rRNA gene sequence similarity to the type strains of Planktotalea lamellibrachiae (97.3%), Halocynthiibacter namhaensis (97.2%), Pseudohalocynthiibacter aestuariivivens (96.8%) and Halocynthiibacter arcticus (96.4%) in the family Rhodobacteraceae. The predominant fatty acids were C10 : 0 3-OH and summed feature 8 (comprising C18 : 1 ω7c and/or C18 : 1 ω6c). The major polar lipids were phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, one unidentified aminolipid and three unidentified lipids. Q-10 was the respiratory quinone. Strain M26A2MT exerted significant algicidal activity against C. polykrikoides cells by destroying the membrane integrity and the photosynthetic system. Our findings suggest that strain M26A2MT shows a high potential to control outbreaks of C. polykrikoides. Based on the polyphasic characterization, strain M26A2MT is considered to represent a novel species within a novel genus of the family Rhodobacteraceae, for which the name Cochlodiniinecator piscidefendens gen. nov., sp. nov. is proposed. The type strain is M26A2MT (=KCTC 82083T=JCM 34119T).

Mariniflexile maritimum sp. nov., isolated from seawater of the South Sea in the Republic of Korea.

A novel Gram-stain-negative, rod-shaped, aerobic, non-motile bacterial strain, designated M5A1MT, was isolated from seawater collected from the South Sea of the Republic of Korea. Based on 16S rRNA gene sequence similarity, strain M5A1MT was closely related to Mariniflexile gromovii KMM 6038T (95.3 %), Mariniflexile fucanivorans SW5T (95.2 %), Mariniflexile soesokkakense RSSK-9T (95.1 %), Yeosuana aromativorans GW1-1T (94.6 %) and Confluentibacter lentus HJM-3T (94.6 %). Genome-based phylogenetic analyses revealed that strain M5A1MT formed a distinct cluster with the type strains of the genus Mariniflexile. The major cellular fatty acid constituents (>5 % of the total fatty acids) were iso-C15:0, anteiso-C15 : 0, iso-C15 : 0 3-OH, iso-C15 : 1 G, iso-C16:03-OH and iso-C17 : 0 3-OH. The respiratory quinone was identified as MK-6. The major polar lipids were phosphatidylethanolamine and one unidentified polar lipid. The genomic DNA G+C content of strain M5A1MT was determined to be 37.7 mol%. On the basis of its phenotypic, phylogenetic and chemotaxonomic characteristics, strain M5A1MT is considered to represent a novel species within the genus Mariniflexile, for which the name Mariniflexile maritimum sp. nov. is proposed. The type strain is M5A1MT (=KCTC 72895T=JCM 33982T).

Aquariibacter albus gen. nov., sp. nov., a new member of the order Burkholderiales, isolated from a freshwater aquarium.

A novel bacterium, strain SJAQ100T, was isolated from a freshwater aquarium and was characterized taxonomically and phylogenetically. Strain SJAQ100T was a Gram-stain-negative, aerobic, rod-shaped and non-motile bacterium. The strain grew optimally with 0 % NaCl and at 25-37 °C on Reasoner's 2A agar. Phylogenetic analysis based on the 16S rRNA gene sequences revealed that the strain SJAQ100T clustered with members of Burkholderiales incertae sedis in the order Burkholderiales, but sequence similarities to known species were less than 96.5 %. The genomic DNA G+C content of strain SJAQ100T was 71.2 mol%. Genomic comparisons of strain SJAQ100T with species in the order Burkholderiales were made using the Genome-to-Genome Distance Calculator, average nucleotide identity and average amino acid identity analyses (values indicated ≤22.1, ≤78.1, and ≤68.1 % respectively). Strain SJAQ100T contained C16 : 0 and C16 : 1 ω7c/C16 : 1 ω6c as major fatty acids and Q-8 as the major quinone. The major polyamines were putrescine and cadaverine. Strain SJAQ100T contained phosphatidylethanolamine and diphosphatidylglycerol as major polar lipids. Based on the genotypic, chemotaxonomic and phenotypic results, strain SJAQ100T represents a novel genus and species, Aquariibacter albus gen. nov., sp. nov., which belongs to order Burkholderiales and the class Betaproteobacteria. The type strain is SJAQ100T (=KCTC 72203T=CGMCC 1.18869T=MCC 4385T).

Novosphingobium aquimarinum sp. nov., isolated from seawater.

A novel Gram-stain-negative, aerobic, and rod-shaped bacterial strain, M24A2MT, was isolated from seawater in the Republic of Korea. On the basis of the 16S rRNA gene phylogeny, strain M24A2MT was found to be closely related to Novosphingobium pentaromativorans US6-1T and Novosphingobium mathurense SM117T with pair-wise sequence similarities of 97.4 and 96.9 %, respectively. Phylogenetic analysis of 16S rRNA sequences indicated that M24A2MT formed a branch with Novosphingobium pentaromativorans US6-1T and represented a member of the genus Novosphingobium. The predominant cellular fatty acids were C14 : 0 2-OH, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), and summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c). The polar lipids of strain M24A2MT consisted mainly of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, one unidentified phospholipid, and two unidentified lipids. The respiratory quinone was ubiquinone Q-10. The genomic DNA G+C content was 63.9 %. Given the phenotypic characteristics along with the phylogenetic distinctness and chemotaxonomic features, strain M24A2MT is considered to represent a novel species within the genus Novosphingobium, for which the name Novosphingobium aquimarinum sp. nov. is proposed. The type strain of Novosphingobium aquimarinum sp. nov. is M24A2MT (=KCTC 72894T=JCM 33983T).

Tabrizicola algicola sp. nov. isolated from culture of microalga Ettlia sp.

A novel Gram-stain-negative, aerobic, non-spore-forming, non-motile, and rod-shaped bacterium, strain ETT8T was isolated from a chemostat culture of microalga Ettlia sp. YC001. Optimal growth was with 0-2% NaCl and at 25-37 °C on R2A medium. Phylogenetic analysis based on the 16S rRNA gene and genome sequence showed that strain ETT8T belongs to the genus Tabrizicola, with the close neighbours being T. sediminis DRYC-M-16T (98.1 %), T. alkalilacus DJCT (97.6 %), T. fusiformis SY72T (96.9 %), T. piscis K13M18T (96.8 %), and T. aquatica RCRI19T (96.5 %). The genomic comparison of strain ETT8T with type species in the genus Tabrizicola was analysed using the genome-to-genome distance calculator (GGDC), average nucleotide identity (ANI), and average amino acid identity (AAI) (values indicated ≤17.7, ≤75.4 and ≤71.9 %, respectively). The genomic DNA G+C content of strain ETT8T was 64.4 %, plus C18 : 1 ω6c and C18 : 0-iso were the major fatty acids and Q-10 the major respiratory quinone. Strain ETT8T contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine aminolipid, and four unidentified lipids as the major polar lipids. Based on the chemotaxonomic, genotypic, and phenotype results, strain ETT8T was recognized as a novel species of the genus Tabrizicola for which the name Tabrizicola algicola sp. nov. is proposed. The type strain is ETT8T (=KCTC 72206T=JCM 31893T=MCC 4339T).

Lysobacter profundi sp. nov., isolated from freshwater sediment and reclassification of Lysobacter panaciterrae as Luteimonas panaciterrae comb. nov.

A polyphasic taxonomic study was carried out on strains CHu50b-3-2T and CHu40b-3-1 isolated from a 67 cm-long sediment core collected from the Daechung Reservoir at a water depth of 17 m, Daejeon, Republic of Korea. The cells of the strains were Gram-stain-negative, non-spore-forming, non-motile and rod-shaped. Comparative 16S rRNA gene sequence studies showed a clear affiliation of two strains with γ-Proteobacteria, which showed the highest pairwise sequence similarities to Lysobacter hankyongensis KTce-2T (96.5 %), Lysobacter pocheonensis Gsoil193T (96.3 %), Lysobacter ginsengisoli Gsoil 357T (96.1 %), Lysobacter solanacearum T20R-70T (96.1 %), Lysobacter brunescens KCTC 12130T (95.4 %) and Lysobacter capsici YC5194T (95.3 %). The phylogenetic analysis based on 16S rRNA gene sequences showed that the strains formed a clear phylogenetic lineage with the genus Lysobacter. The major fatty acids were identified as summed feature 9 (iso-C17 : 1 ω9c and/or C18 : 1 10-methyl), iso-C15 : 0, iso-C16 : 0 and iso-C17 : 0. The respiratory quinone was identified as ubiquinone Q-8. The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid. The genomic DNA G+C content was determined to be 66.8 mol% (genome) for strain CHu50b-3-2T and 66.4 mol% (HPLC) for strain CHu40b-3-1. Based on the combined genotypic and phenotypic data, we propose that strains CHu50b-3-2T and CHu40b-3-1 represent a novel species of the genus Lysobacter, for which the name Lysobacter profundi sp. nov. is proposed. The type strain is CHu50b-3-2T (=KCTC 72973T=CCTCC AB 2019129T). Besides Lysobacter panaciterrae Gsoil 068T formed a phylogenetic group together with strain Luteimonas aquatica RIB1-20T (EF626688) that is clearly separated from all other known Lysobacter strains. Based on the phylogenetic relationships together with fatty acid compositions, Lysobacter panaciterrae Gsoil 068T should be reclassified as a member of the genus Luteimonas: Luteimonas aquatica comb. nov. (type strain Gsoil 068T=KCTC 12601T=DSM 17927T).

Paraconexibacter algicola gen. nov., sp. nov., a novel actinobacterium isolated from a eutrophic lake during the end of cyanobacterial harmful algal blooms, and proposal of Paraconexibacteraceae fam. nov. in the order Solirubrobacterales.

A novel bacterium, strain Seoho-28T, was isolated from a shallow eutrophic lake during the end of cyanobacterial harmful algal blooms and was characterized taxonomically and phylogenetically. Strain Seoho-28T was a Gram-stain-negative, aerobic, rod-shaped and non-motile bacterium. The strain grew optimally with 0 % NaCl and at 25-30 °C on Reasoner's 2A medium. The phylogenetic analysis based on 16S rRNA gene sequences positioned the novel strain among the order Solirubrobacterales, but sequence similarities to known species were less than 94.7 %. The genomic DNA G+C content of the strain Seoho-28T was 74.2 mol%. Genomic comparisons of strain Seoho-28T with families in the order Solirubrobacterales were made using the Genome-to-Genome Distance Calculator, average nucleotide identity and average amino acid identity analyses (values indicated ≤14.9, ≤73.5 and ≤57.8 %, respectively). Strain Seoho-28T contained C16 : 0-iso, C18 : 1 ω9c and C16 : 0 as major fatty acids and MK-7 (H4) as the major quinone. Strain Seoho-28T contained diphosphatidylglycerol, phosphatidylinositol and an unidentified phospholipid as major polar lipids. Meso- and ll-diaminopimelic acids were the diagnostic diamino acids in the cell-wall peptidoglycan. Based on the genotypic, chemotaxonomic and phenotypic results, strain Seoho-28T represents a novel genus and species, Paraconexibacter algicola gen. nov., sp. nov., which belongs to a new family Paraconexibacteraceae in the order Solirubrobacterales and the class Thermoleophilia. The type strain is Seoho-28T (=KCTC 39791T=JCM 31881T).

Genomic insights into a novel species Rhodoferax aquaticus sp. nov., isolated from freshwater.

A novel non-phototrophic member of the genus Rhodoferax was obtained from freshwater. The purpose of this study was to analyse the genome of a nonphototrophic strain and propose a new species based on its phylogenetic, genomic, physiological and chemotaxonomic characteristics. The results of phylogenetic analysis based on 16S rRNA gene sequences supports that the strain, designated Gr-4T, has a close relationship to the genus Rhodoferax. The observed average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain Gr-4T and its closest related strains were 72.3-74.6 % and 21.9-22.8 %, respectively. These values were much lower than the species separation thresholds for ANI or dDDH of 95-96 and 70 %, respectively, and in fact fall in the intergeneric range. Strain Gr-4T does not contain RuBisCO-related genes, but does contain GS/GOGAT pathway-related genes enabling nitrate ammonification. A polyphasic study and a genomic-level investigation were done to establish the taxonomic status of strain Gr-4T. Based on the phylogenetic, genomic and physiological differences, it is proposed that the isolate be classified to the genus Rhodoferax as Rhodoferax aquaticus sp. nov. with isolate Gr-4T (=KCTC 32394T=JCM 19166T) as the type strain.

Lacisediminimonas profundi gen. nov., sp. nov., a member of the family Oxalobacteraceae isolated from freshwater sediment.

A novel Gram-stain-negative bacterial strain, CHu64-6-4T, was isolated from a 67-cm-long sediment core collected from the Daechung Reservoir at a water depth of 17 m, Daejeon, Republic of Korea. The cells of strain CHu64-6-4T were aerobic nonmotile and formed colorless colonies on R2A agar. The phylogenetic analysis based on 16S rRNA gene sequencing indicated that the strain formed a separate lineage within the family Oxalobacteraceae, exhibiting 97.2% and 97.1% 16S rRNA gene sequence similarities to Glaciimonas singularis and Paraherbaspirillum soli, respectively. Strain CHu64-6-4T showed less than 74.4% average nucleotide identity compared to the type strains of related genera within the family Oxalobacteraceae. In the UPGMA dendrogram based on the ANI values of genomic sequences, strain CHu64-6-4T formed an evolutionary lineage independent of the genera Glaciimonas and some other taxa. The chemotaxonomic results showed Q-8 as the predominant respiratory ubiquinone, phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylethnolamine as the major polar lipids, Summed Feature 3 (C16:1ω7c and/or iso-C15:0 2-OH), C16:0, and C18:1ω7c as the major fatty acids, and a DNA G+C content of 62.1 mol%. The combined genotypic and phenotypic data showed that strain CHu64-6-4T could be distinguished from all genera within the family Oxalobacteraceae and represents a novel genus, Lacisediminimonas profundi gen. nov., with the name Lacisediminimonas profundi sp. nov., in the family Oxalobacteraceae. The type strain is CHu64-6-4T (=KCTC 62287T=JCM 32676T).